Waters Alliance LC/MS System Analytical Conditions Chromatograms Mass Spectra Calibration Quantitation Analysis of Pesticides (II) Metribuzin & their metabolites in Rice Jun Yonekubo,, JAPAN Key Words Pesticides, sensitivity, calibration, electrospray 1
Analytical Conditions Waters Alliance 269 HPLC Pump Conditions Column Symmetry C18, 3.5 um, 2.1 x 15 mm.3 AcOH aq. 75. ACN 18. MeOH 7. Flow (ml/min).3 Run Time (mins) 15. Column Temperature ( C) 4. Waters ZMD MS Detector Conditions MS Ion Energy: 3.5 Volts LM Resolution: 1.7 HM Resolution: 1.7 Multiplier: 65 Volts Pressures Analyser Vacuum: 2.6e-5 mbar Gas Flow: 3.4 lit/hr Ionization mode: ES/APcI+ Function type: Scan Mass range: 15 to 25 Cone Voltage: 35 Ionization mode: ES/APcI- Function type: Scan Mass range: 15 to 25 Cone Voltage: 35 2
Metribuzin and their 3 metabolites (5ppm) standard mixture 5.24 21 6.39 21 12.48 21 8.98 238 4 mix, 5ppm 144 (5.287) 185. DK:MW=184 1: Scan ES+ 4.53e5 3: Diode Array 28 8.7e3-7 9.3 2 12.51 215 157.1 226. 175 (6.442) 168. 2: Scan ES- 1.31e5 DADK:MW=169 1: Scan ES+ 1.13e6 5.29 185 246 (9.27) 2. 1: Scan ES+ 6.38e5 DA:MW=199 15 9.1 198 172. 341 (12.511) 215. 1: Scan ES+ 7.97e5 2: Scan ES- 4.44e5 5.31 183 6.44 168 Metribuzin:MW=214 187. 15 16 17 18 19 2 21 22 23 24 25 m/z 1 4. 6. 8. 1. 12. 14. 16. 18. 2. 22. 24. 26. 28. 3. 32. 34. Time 3
ESI (left) and APcI (right) detection of 5ppm std.mixture. 5.24 21 6.39 21 12.48 21 3: Diode Array 28 8.26e3 3: Diode Array 28-2.5e4 8.98 238-53 ESI positive 5.29 185 9.6 2 12.51 215 151 1: Scan ES+ 1.17e6 APcI positive 8.99 2 12.51 215 1: Scan AP+ 4.3e4 5.29 185 15 ESI negative 9.1 198 3 2: Scan ES- 4.71e5 2: Scan AP- 3.14 5.235.56 152 6.59 9.41 152 7.73;152 9.741.7 152 12.2414.11 152 152 152 152 3.39e6 152 152 APcI negative 5.31 183 6.44 168 1 4. 6. 8. 1. 12. 14. Time 43 4. 6. 8. 1. 12. 14. Time 4
ESI (left) and APcI (right) detection of.5ppm standard mixture 5_Metribuzin mix 5.28 53897 ESI 1: SIR of 1 Channel ES- 183. 2.11e6 5.31 11882 1: SIR of 1 Channel AP+ 185. 4.89e4 APcI 5_Metribuzin mix 6.42 48931 6 2: SIR of 1 Channel ES- 168. 1.78e6 6.42 182994 5.3 8692 2: SIR of 1 Channel AP- 168. 1.16e6 5_Metribuzin mix 8.99 199286 16 3: SIR of 1 Channel ES- 198. 3.35e6 9.2 68535 3: SIR of 1 Channel AP+ 2. 2.22e5 5_Metribuzin mix 4: SIR of 1 Channel ES+ 3 12.52 215. 277196 7.77e6 4: SIR of 1 Channel AP+ 12.58 215. 58365 1.73e5 4 2. 4. 6. 8. 1. 12. 14. Time 15 2. 4. 6. 8. 1. 12. 14. Time 5
1ppb standard mixture (left) and Spiked Rice Sample (right) 1_Metribuzin mix 5.28 19 1: SIR of 1 Channel ES- 183. 3.93e4 1_spike 5.28 11283 1: SIR of 1 Channel ES- 183. 4.5e4 5 1_Metribuzin mix 6.38 9894 2: SIR of 1 Channel ES- 168. 3.39e4 1_spike 6 6.39 1728 2: SIR of 1 Channel ES- 168. 3.76e4 12 1_Metribuzin mix 8.97 28446 3: SIR of 1 Channel ES- 198. 8.12e4 12 1_spike 8.95 3288 3: SIR of 1 Channel ES- 198. 8.68e4 2 1_Metribuzin mix 4: SIR of 1 Channel ES+ 12.47 215. 62839 4.77e5 3 1_spike 4: SIR of 1 Channel ES+ 12.51 215. 65788 6.88e5 69 5. 1. Time 57 5. 1. Time 6
Compound 1 name: DK Coefficient of Determination:.996118 Calibration curve: 97567 * x + Response type: External Std, Curve type: Linear, Origin: Force, Weighting: Null, Axis trans: None 9.76e5 Response..1.2.3.4.5.6.7.8.9 1. Conc Compound 2 name: DADK Coefficient of Determination:.992975 Calibration curve: 874956 * x + Response type: External Std, Curve type: Linear, Origin: Force, Weighting: Null, Axis trans: None 8.75e5 Response Conc..1.2.3.4.5.6.7.8.9 1. 7
Compound 3 name: DA Coefficient of Determination:.999 Calibration curve: 1.97741e6 * x + Response type: External Std, Curve type: Linear, Origin: Force, Weighting: Null, Axis trans: None 1.98e6 Response Conc..1.2.3.4.5.6.7.8.9 1. Compound 4 name: Metribuzin Coefficient of Determination:.997558 Calibration curve: 5.25398e6 * x + Response type: External Std, Curve type: Linear, Origin: Force, Weighting: Null, Axis trans: None 5.25e6 Response Conc..1.2.3.4.5.6.7.8.9 1. 8
Quantify Compound Summary Report (I ) Compound 1: DK Name Type Std Conc* RT Conc.* BS/Conc.* 1_Metribuzin mix Standard.1 5.279 19.1.1 5_Metribuzin mix Standard.5 5.28 529.5.5 1_Metribuzin mix Standard.1 5.28 1986.11.11 5_Metribuzin mix Standard.5 5.28 53897.54.54 1_Metribuzin mix Standard 1. 5.246 952662.98.98 Control Analyte 5.8 448.. 1_spike Analyte 5.28 11283.1.1 5_spike Analyte 5.279 5661.6.6 Compound 2: DADK Name Type Std Conc* RT Conc.* BS/Conc.* 1_Metribuzin mix Standard.1 6.385 9894.1.1 5_Metribuzin mix Standard.5 6.419 48219.6.6 1_Metribuzin mix Standard.1 6.419 12653.12.12 5_Metribuzin mix Standard.5 6.419 48931.56.56 1_Metribuzin mix Standard 1. 6.385 847429.97.97 Control Analyte 6.216 593.. 1_spike Analyte 6.386 1728.1.1 5_spike Analyte 6.418 56317.6.6 * ppm 9
Quantify Compound Summary Report (II ) Compound 3: DA Name Type Std Conc* RT Conc.* BS/Conc.* 1_Metribuzin mix Standard.1 8.969 28446.1.1 5_Metribuzin mix Standard.5 9.21 13493.7.7 1_Metribuzin mix Standard.1 9.4 273264.14.14 5_Metribuzin mix Standard.5 8.986 199286.56.56 1_Metribuzin mix Standard 1. 8.953 1912679.97.97 Control Analyte 8.767 1797.. 1_spike Analyte 8.953 3288.2.1 5_spike Analyte 8.986 146491.7.7 Compound 4: Metribuzin Name Type Std Conc* RT Conc.* BS/Conc.* 1_Metribuzin mix Standard.1 12.471 62839.1.1 5_Metribuzin mix Standard.5 12.49 324447.6.6 1_Metribuzin mix Standard.1 12.474 642414.12.12 5_Metribuzin mix Standard.5 12.524 277196.53.53 1_Metribuzin mix Standard 1. 12.44 51673.98.98 Control Analyte 12.338 22315.. 1_spike Analyte 12.57 65788.1.1 5_spike Analyte 12.489 342316.7.7 * ppm 1